Membrane transport and cytokinin action in root hairs of Medicago sativa

In an effort to develop a cellular model for studying cytokinin action in higher plants, we investigated the effect of cytokinins on growth and membrane transport in root hairs of alfalfa (Medicago sativa L.) seedlings. Alfalfa seedlings grown for 24 h in the presence of cytokinins showed increased root hair length and formation of root hairs close to the root cap. Increased growth of root hairs was observed within 10 min of cytokinin application and was accompanied by a hyperpolarization of the plasma membrane. The ion-transport inhibitor diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) blocked both cytokinin-induced root hair growth and hyperpolarization but did not, by itself, alter growth or membrane potential. Finally, hyperpolarization was induced by extracellular cytokinin but not by injection of cytokinin into the cytoplasm. These findings show that root hairs undergo several rapid responses to cytokinin, including changes in membrane transport and growth. We conclude that multiple cytokinin response in root hairs may be mediated by events involving perception and ion transport at the plasma membrane.